Somatic Mutation Identification and Verification

Tumor heterogeneity, sample source, sample preparation — all of these factors can affect the ratio of mutated to wildtype DNA, rendering somatic mutations nearly undetectable. For samples in which somatic mutations may be vanishingly scarce, such as FFPE, serum/plasma, and LCM samples, preamplification holds the key to rescuing mutation detection. Read on to find out more about QIAGEN's somatic mutation detection solutions. ...

Read more

Tumor heterogeneity, sample source, sample preparation — all of these factors can affect the ratio of mutated to wildtype DNA, rendering somatic mutations nearly undetectable. For samples in which somatic mutations may be vanishingly scarce, such as FFPE, serum/plasma, and LCM samples, preamplification holds the key to rescuing mutation detection. Read on to find out more about QIAGEN's somatic mutation detection solutions.

QIAGEN has developed a robust somatic mutation detection assay pipeline that covers over 1200 cancer-related mutations in over 150 genes. qBiomarker Somatic Mutation PCR Arrays are panels of mutation-specific real-time PCR assays for simultaneously profiling mutations within a disease- or pathway-focused set of genes. These assays and arrays provide users a bench-verified solution for the verification or focused profiling of mutations in cancer samples by real-time PCR.

Cancer-causing mutations have been identified and cataloged in databases such as COSMIC. Within these databases are mutations ranging from simple nucleotide substitutions to small deletions and insertions. These mutations are spread across the genome, from the cancer gene-rich chromosome 17 to the cancer gene-poor chromosome 20.

Somatic mutations are challenging to detect because of tumor and sample heterogeneity. Samples comprise both tumor and wildtype tissue, which can leave the investigator with a “diluted” sample. Further complicating detection is the fact that some tumor cells acquire new mutations that can change the cellular phenotype. The new qBiomarker PreAMP Synthesis Kit and qBiomarker PreAMP Mixes preamplify DNA from samples with limited amplifiable DNA to enhance sensitivity. qBiomarker Somatic Mutation PCR Assays preferentially detect the mutant allele, generating signal from a small amount of template while not amplifying the wildtype allele that would generate background signal.

Mutation assays can then be grouped together into PCR arrays based off common themes to enable users to profile a focused set of mutations. The mutations on an array are selected based on a commonality, such as being from the same gene, signaling pathway, or cancer type. For example, users could profile the most common 84 mutations in breast cancer or the 44 most common mutations in APC. The Custom qBiomarker Somatic Mutation PCR Arrays allow users to select the mutations that they wish to profile. Together, the qBiomarker Somatic Mutation products facilitate sensitive detection and profiling of mutations in cancer cells or tumor samples.